Open AccessEffect of N-cadherin on Chondrogenic Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells through Wnt Signaling Pathway
Author(s) -
Feng Qu,
Xueqin Shen,
Haipeng Li,
Jingyang Zhou,
Biao Yuan,
Chunbao Li,
Wei Qi,
Yujie Liu,
Mingzhu Zhang
Publication year - 2022
Publication title -
cellular and molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.371
H-Index - 71
eISSN - 1165-158X
pISSN - 0145-5680
DOI - 10.14715/cmb/2021.67.6.33
Subject(s) - wnt signaling pathway , mesenchymal stem cell , cadherin , chondrogenesis , western blot , chemistry , microbiology and biotechnology , stem cell , catenin , sox9 , staining , bone morphogenetic protein 2 , regeneration (biology) , bone marrow , biology , signal transduction , pathology , immunology , medicine , gene expression , cell , in vitro , biochemistry , gene
This study aimed to compare and analyze the effect of N-cadherin on chondrogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) and to explore the related mechanism, so as to provide a novel theoretical basis for the clinical work of articular cartilage injury regeneration and repair. For this purpose, the experimental animals were clean grade SD rats (aged 5-6 weeks, weighing 180-250g). Alcian blue staining was carried out to observe the induced chondrogenesis following N-cadherin inhibition. The specific role of N-cadherin in the Wnt signaling pathway and chondrogenic differentiation of BMSCs was detected by Western blot; while the effect of N-cadherin on the molecular level changes of β-catenin in the cytoplasm was evaluated by fluorescence quantitative real-time PCR (qRT-PCR). In addition, immunoprecipitation (IP) was used for the verification of the interaction between N-cadherin and β-catenin. Results showed that under the light microscope, 90% of the BMSCs at the third generation, 90% of the cells were fused. Alcian blue staining showed that the green staining area in the BMP2 induction group was large and dense, while that in the N-cadherin inhibition group and blank control group was small and sparse. Western blot revealed that N-cadherin and SOX9 were significantly developed in the BMP2 induction group, but Wnt3a was not significantly developed. While in the N-cadherin inhibition group, the development of Wnt3a was obvious, yet without evident development of N-cadherin and SOX9. The qRT-PCR indicated that the relative mRNA expression of Wnt3a was significantly increased in the N-cadherin inhibition group (P 0.05). Western blot indicated that in the BMP2 induction group; there existed the development of β-catenin, significant development of phos-GSK-3β and total GSK-3β, but no obvious development of Wnt3a. In the N-cadherin inhibition group, there was significantly enhanced development of Wnt3a and β-catenin than that before, blurred development of phos-GSK-3β than that before, and also obvious development of total GSK-3β with little change from before. N-cadherin promoted the expression of β-catenin mostly in the cell membrane, but only a few in the cytoplasm and nucleus. Additionally, verification by IP showed that N-cadherin and β-catenin were developed on N-cadherin and β-catenin bands, suggesting an interaction between N-cadherin and β-catenin. According to these results, N-cadherin can ultimately promote chondrogenic differentiation of BMSCs by inhibiting the Wnt signaling pathway.